Research Article Open Access

Detection and Localization of Anti and Pro-apoptotic mRNA Genes in Human Colorectal Cancer Using in situ RT-PCR

Mohd. Nazil Salleh1, Boo Siaw Shi1, Wan Khairuzzaman Wan Ramli1, Azlina Zahari1, Norashikin Shamsudin2 and Thuaibah Hashim3
  • 1 University Industry Selangor, Malaysia
  • 2 University Putra Malaysia, Malaysia
  • 3 Cyberjaya University College of Medical Sciences, Malaysia

Abstract

Problem statement: Recent studies using conventional Polymerase Chain Reaction have shown that anti-apoptotic (Cyclooxygenase-2, COX-2 and Nuclear Factor-κB, NF-κB) and proapoptotic mRNA (Bax and Bad) are involved in the tumorigenesis of colorectal cancer. The aim of this study was to localize the expression of anti and pro-apoptotic mRNA genes using Reverse Transcription in situ Polymerase Chain Reaction (RT in situ PCR) and immunodetection technique in the early stage of human colorectal adenocarcinoma. Approach: Reverse Transcription in situ Polymerase Chain Reaction (RT in situ PCR) and immunodetection technique was applied throughout of this studies. 20 paraffin-embedded tissue blocks of human colorectal adenocarcinoma samples was used compared to controls. Results: Morphologically, the glands and crypts were well-differentiated, enlarged and irregular with active secretion of mucin. COX-2, NF-κB, Bax and Bad mRNA were expressed in both normal and human colorectal cancer tissues. All mRNA genes were expressed in the cytoplasm and nuclei. However, COX-2 and NF-κB mRNA genes were highly expressed with higher intensity of brown staining compared to Bax and Bad at tubular epithelium cells. Conclusion: This study demonstrated that by using RT in situ PCR, COX-2 and NF-κB mRNA genes were shown to be involved in the development of human colorectal cancer.

American Journal of Biochemistry and Biotechnology
Volume 6 No. 3, 2010, 164-171

DOI: https://doi.org/10.3844/ajbbsp.2010.164.171

Submitted On: 28 July 2010 Published On: 30 September 2010

How to Cite: Salleh, M. N., Shi, B. S., Ramli, W. K. W., Zahari, A., Shamsudin, N. & Hashim, T. (2010). Detection and Localization of Anti and Pro-apoptotic mRNA Genes in Human Colorectal Cancer Using in situ RT-PCR. American Journal of Biochemistry and Biotechnology, 6(3), 164-171. https://doi.org/10.3844/ajbbsp.2010.164.171

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Keywords

  • Cyclooxygenase-2 (COX-2)
  • Nuclear Factor-κB (NF-κB)
  • heterodimeric partner for Bcl- XL and Bcl-2 (Bad)
  • bcl-2 associated X protein (Bax)
  • in situ Reverse Transcription Polymerase Chain Reaction (in situ RT-PCR)